人参皂苷Rb1、Rg1对胎鼠体外神经干细胞促增殖分化作用的研究

关键词：神经干细胞  体外培养  人参皂苷Rb1、Rg1  增殖  分化

Keywords:neural stem cells  in vitro culture  ginsenoside Rb1, Rg1  proliferation  differentiation

摘要：

      摘要 目的：探索人参皂苷Rb1、Rg1不同浓度对体外培养神经干细胞（neural stem cells，NSCs）增殖和分化的影响。 方法：对胎鼠大脑皮质源性NSCs进行体外培养、鉴定；用5-溴脱氧尿嘧啶核苷（BrdU）掺入法测定不同浓度的人参皂苷Rb1、Rg1及对照组（空白组）对NSCs增殖的影响，通过计算BrdU/DAPI比值，确定其促增殖的适宜浓度；荧光免疫细胞化学技术检测各组Tuj1、GFAP及DAPI（4′, 6-diamidino-2-phenylindole）的表达，计算Tuj1/DAPI、GFAP/DAPI百分比，比较人参皂苷Rb1、Rg1对NSCs分化为神经元和神经胶质细胞的效率。 结果：①所培养的细胞经鉴定为NSCs后，增殖实验中除人参皂苷0.1μmol/L Rg1（P＞0.05）外，其余各组Rg1和Rb1的BrdU/DAPI比值均显著性增加（P＜0.05），Rb1和Rg1均能促进体外培养的NSCs增殖，其适宜浓度Rb1、Rg1分别为1μmol/L、10μmol/L。②人参皂苷Rb1（10μmol/L）组GFAP/DAPI百分比与对照组相比显著增高（P＜0.05），而人参皂苷Rg1组GFAP/DAPI百分比及两种人参皂苷Tuj1/DAPI比值与对照组相比无显著性差异（P＞0.05）。 结论：人参皂苷Rg1、Rb1在一定浓度范围内可促进体外NSCs增殖。Rb1能促进NSCs分化为星形胶质细胞。

Abstract:

      Abstract Objective: To investigate the effects of ginsenoside Rb1, Rg1 with different concentrations on the proliferation and differentiation of NSCs in vitro. Method: NSCs were isolated from the cerebral cortex of embryonic mice. The cells were cultured in vitro and identified by immunocytochemistry staining. The ginsenoside Rb1, Rg1 were co-incubated with NSCs at 0.1 μM, 1μM and 10μM respectively for 48 hours. The BrdU/DAPI ratio was calculated to detect the effect of ginsenosides on the proliferation of NSCs, which identified the most effective concentration for NSCs proliferation. After incubation with 10μM Rb1, for 5—7 days, the differentiation of NSCs into neurons and glial cells was determined by calculating the Tuj1/DAPI and GFAP/DAPI ratio using immunofluorescence staining. Result: ①After the cultured cells were identified as neural stem cells,we performed proliferation experiment. We found that all ginsenosides obviously promoted the proliferation of NSCs (P＜0.05) except a concentration of 0.1μmol/L Rg1（P＞0.05）. ② The ginsenoside Rb1 promoted astrocytic differentiation compared with the control group(P＜0.05), while Rg1 had no effect on enhancing astrocytic differentiation (P＞0.05). No effects on neuronal differentiation of NSCs were found in two ginsenosides treated groups. Conclusion: Ginsenoside Rb1, Rg1 could promote the proliferation of NSCs in vitro at certain concentrations. Rb1 could promote astrocytic differentiation of NSCs.